Effectiveness of Lactobacillus acidophilus 2B4 as Biocontrol to Prevent Salmonella enteritidis Infection on Laying Hens
Effectiveness of Lactobacillus acidophilus 2B4 as Biocontrol to
Prevent Salmonella enteritidis Infection on Laying Hens
Niken Ulupi1, Irma Isnafia Arief1,*, Bram Brahmantiyo2 & Kharisma Eka Riwayati1
1
Department of Animal Production and Technology, Faculty of Animal Science,
Bogor Agricultural University, Bogor, 16680, Indonesia,
*Email: irma_isnafia@yahoo.com
2
Indonesian Research Institute for Animal Production, Bogor,
Jl.Veteran III, Banjarwaru PO Box 221, Bogor 16002, Indonesia
Abstract
Laying hen is raised and selected to produce eggs. Poultry could be infected
by several kinds of Salmonella enterica such as S. enteritidis as a specific bacterium
which is carried by chickens. Salmonella enteritidis can depress the hen weight by
dehydration and contaminate the egg which is very dangerous for human health.
Regarding this negative effect, the preventive treatment to eliminate Salmonella
enteritidis contamination in eggs becomes a major concern. This research aimed
to determine the optimum dosage and frequency applied of probiotic L.acidophilus
to avoid contamination of Salmonella enteritidis in ovary and egg of laying hens.
The result showed that probiotic treatment could increase feed consumption, egg
production, however, in the other hand decreased the feed conversion. Among other,
probiotic treatment was effective to reduce the population of Salmonella enteritidis
in ovary and egg of laying hens. The optimum dosage of L.acidophilus probiotic was
within population of 109 cfu/ml and frequency of twice a day was the most effective
as biocontrol.
Key words: L. acidophilus, laying hen, probiotic, Salmonella enteritidis
Introducton
Food securty s a major ssue n natonal development program. Food s a basc
requrement for the fulfllment of - human rghts for everyone. Food safety ssues
are major concern n publc health polcy. Foodborne dsease and food contamnaton
ncdents occurred n varous countres, not only n developng countres where
santaton and hygene condtons are generally poor, but also n developed countres.
One of the emergng pathogen s Salmonella enteritidis transovaran contamnaton
of grade A eggs. These eggs are contamnated snce the start of ts formaton n the
body because ts parent s infected by S.enteritidis n ovares (Gantos et al., 2009).
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
689
Bosecurty va bocontrol s needed to apply n layer chcken farms as preventve
effort.
Lactobacillus acidophilus 2B4 has been proven as probotc (Aref, 2011). Ths
stran could nhbt the growth of S. enteritidis solated from the ovary of layng hen
by n vtro analyss (Ulup et al., 2009). Applcaton of probotc L. acidophilus 2B4
as a bocontrol agent for the preventon of contamnaton of eggs by S. enteritidis
transovaran s very necessary, especally n layng hens. The am of ths research
was to fnd out the dosage and frequency of probotc L. acidophilus 2B4 treatment
to prevent contamnaton of S. enteritidis n ovarum and egg of layer hen
Materals and Methods
Sxty layer hens were dvded nto 4 Treatments. Treatment 1 (R1) was
layng hens wthout oral admnstraton of L. acidophilus 2B4 and S. enteritidis
(as control). R2 was layng hens wthout L. acidophilus 2B4, but were nfected
by S. enteritidis. R3 was layng hens admnstered of L. acidophilus 2B4 once per
day (dosage 1 x109 cfu/ml) and were nfected by S. enteritidis. R4 was layng hens
wth oral admnstraton twce per day (dosage 2 x109 cfu/ml) and were nfected by
S. enteritidis. L. acidophilus 2B4 was gven va drnkng water for 20 days, and S.
enteritidis was gven by oral admnstraton for 5 days (day 5-day 9). Each treatment
conssted of 15 layng hens. , A total of 3 hens of each treatment were slaughtered
for parameters evaluaton. n day 5, 10, 15 and 20.
Parameters observed were performances of layer hens and Salmonella contamnaton on ovarum and eggs. Analyss of Salmonella contamnaton was done
by qualtatve analyss accordng to BAM (2007). The samples were ncubated on
selectve meda on Lysne Desoxycholate xylose meda (XLD) Agar, Hectoen Eterc
Agar (HEA), and Bsmuth sulfte agar (BSA). These three selectve meda were
ncubated at 35 ± 2 °C for 24 ± 2 hours. After ncubaton typcally appearance was
observed whether there was a growng colony. The analyss followed the bochemcal tests usng trple sugar early Iron (TSI) and Lysne Iron Agar (LIA) n talcs.
Typcal colones that grew on the three specfc medas XLD Agar, HE agar and BS.
Each loop was noculated usng a sterle needle on TSI agar and LIA agar.
The expermental was completely random desgned wth 3 replcatons. Data
were analyzed usng ANOVA and then further subjected to Tukey test (Steel and
Torre, 1995) f there were any dfferences.
Results and Dscusson
Performance of layer hens
Performance of layer hens were affected by probotc admnstraton as
descrbed n Table 1.
690
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
Table 1. Performance of layer hens
Performance
Feed consumpton (g/hen/day)
Egg producton (%)
Egg weght (g/egg)
Feed converson
Mortalty
Haugh Unt (HU)
R1
R2
a
111.81
67.64a
59.43
3.34a
0
96.40(AA)
R3
a
112.80
70.31a
58.89
3.48a
0
94.00(AA)
R4
b
118.29
75.91b
59.57
3.36a
0
94.59(AA)
118.54b
81.97c
59.58
2.72b
0
95.30(AA)
Dfferent superscrpt n the same lne means sgnfcantly dfferent (P
Prevent Salmonella enteritidis Infection on Laying Hens
Niken Ulupi1, Irma Isnafia Arief1,*, Bram Brahmantiyo2 & Kharisma Eka Riwayati1
1
Department of Animal Production and Technology, Faculty of Animal Science,
Bogor Agricultural University, Bogor, 16680, Indonesia,
*Email: irma_isnafia@yahoo.com
2
Indonesian Research Institute for Animal Production, Bogor,
Jl.Veteran III, Banjarwaru PO Box 221, Bogor 16002, Indonesia
Abstract
Laying hen is raised and selected to produce eggs. Poultry could be infected
by several kinds of Salmonella enterica such as S. enteritidis as a specific bacterium
which is carried by chickens. Salmonella enteritidis can depress the hen weight by
dehydration and contaminate the egg which is very dangerous for human health.
Regarding this negative effect, the preventive treatment to eliminate Salmonella
enteritidis contamination in eggs becomes a major concern. This research aimed
to determine the optimum dosage and frequency applied of probiotic L.acidophilus
to avoid contamination of Salmonella enteritidis in ovary and egg of laying hens.
The result showed that probiotic treatment could increase feed consumption, egg
production, however, in the other hand decreased the feed conversion. Among other,
probiotic treatment was effective to reduce the population of Salmonella enteritidis
in ovary and egg of laying hens. The optimum dosage of L.acidophilus probiotic was
within population of 109 cfu/ml and frequency of twice a day was the most effective
as biocontrol.
Key words: L. acidophilus, laying hen, probiotic, Salmonella enteritidis
Introducton
Food securty s a major ssue n natonal development program. Food s a basc
requrement for the fulfllment of - human rghts for everyone. Food safety ssues
are major concern n publc health polcy. Foodborne dsease and food contamnaton
ncdents occurred n varous countres, not only n developng countres where
santaton and hygene condtons are generally poor, but also n developed countres.
One of the emergng pathogen s Salmonella enteritidis transovaran contamnaton
of grade A eggs. These eggs are contamnated snce the start of ts formaton n the
body because ts parent s infected by S.enteritidis n ovares (Gantos et al., 2009).
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
689
Bosecurty va bocontrol s needed to apply n layer chcken farms as preventve
effort.
Lactobacillus acidophilus 2B4 has been proven as probotc (Aref, 2011). Ths
stran could nhbt the growth of S. enteritidis solated from the ovary of layng hen
by n vtro analyss (Ulup et al., 2009). Applcaton of probotc L. acidophilus 2B4
as a bocontrol agent for the preventon of contamnaton of eggs by S. enteritidis
transovaran s very necessary, especally n layng hens. The am of ths research
was to fnd out the dosage and frequency of probotc L. acidophilus 2B4 treatment
to prevent contamnaton of S. enteritidis n ovarum and egg of layer hen
Materals and Methods
Sxty layer hens were dvded nto 4 Treatments. Treatment 1 (R1) was
layng hens wthout oral admnstraton of L. acidophilus 2B4 and S. enteritidis
(as control). R2 was layng hens wthout L. acidophilus 2B4, but were nfected
by S. enteritidis. R3 was layng hens admnstered of L. acidophilus 2B4 once per
day (dosage 1 x109 cfu/ml) and were nfected by S. enteritidis. R4 was layng hens
wth oral admnstraton twce per day (dosage 2 x109 cfu/ml) and were nfected by
S. enteritidis. L. acidophilus 2B4 was gven va drnkng water for 20 days, and S.
enteritidis was gven by oral admnstraton for 5 days (day 5-day 9). Each treatment
conssted of 15 layng hens. , A total of 3 hens of each treatment were slaughtered
for parameters evaluaton. n day 5, 10, 15 and 20.
Parameters observed were performances of layer hens and Salmonella contamnaton on ovarum and eggs. Analyss of Salmonella contamnaton was done
by qualtatve analyss accordng to BAM (2007). The samples were ncubated on
selectve meda on Lysne Desoxycholate xylose meda (XLD) Agar, Hectoen Eterc
Agar (HEA), and Bsmuth sulfte agar (BSA). These three selectve meda were
ncubated at 35 ± 2 °C for 24 ± 2 hours. After ncubaton typcally appearance was
observed whether there was a growng colony. The analyss followed the bochemcal tests usng trple sugar early Iron (TSI) and Lysne Iron Agar (LIA) n talcs.
Typcal colones that grew on the three specfc medas XLD Agar, HE agar and BS.
Each loop was noculated usng a sterle needle on TSI agar and LIA agar.
The expermental was completely random desgned wth 3 replcatons. Data
were analyzed usng ANOVA and then further subjected to Tukey test (Steel and
Torre, 1995) f there were any dfferences.
Results and Dscusson
Performance of layer hens
Performance of layer hens were affected by probotc admnstraton as
descrbed n Table 1.
690
Proceeding of the 2nd International Seminar on Animal Industry | Jakarta, 5-6 July 2012
Table 1. Performance of layer hens
Performance
Feed consumpton (g/hen/day)
Egg producton (%)
Egg weght (g/egg)
Feed converson
Mortalty
Haugh Unt (HU)
R1
R2
a
111.81
67.64a
59.43
3.34a
0
96.40(AA)
R3
a
112.80
70.31a
58.89
3.48a
0
94.00(AA)
R4
b
118.29
75.91b
59.57
3.36a
0
94.59(AA)
118.54b
81.97c
59.58
2.72b
0
95.30(AA)
Dfferent superscrpt n the same lne means sgnfcantly dfferent (P